TLR1/2 agonist treatment is associated with a loss of DCs and macrophages in the bone marrow and HSPC expansion and mobilization. Wild-type mice were treated with PAM₃CSK₄ (100 μg, every other day × 3 doses) and analyzed 24 hours after the final dose. (A) Total leukocytes per femur. (B-C) Number of the indicated cell type in the bone marrow (BM). Number of LSK (D) or LSK-SLAM (E) cells in the BM. Number of LSK (F) and LSK-SLAM (G) cells in the spleen (SP). (H) Number of LSK cells in the peripheral blood (PB). Number of colony-forming cell (CFC) in BM (I), SP (J), or PB (K). (L) Representative flow plots showing gating strategy to identify different stages of the cell cycle; data are gated on LSK cells. Cell cycle analysis of LSK (M) and LSK-SLAM (N) cells in the BM. Data represent the mean ± standard error of the mean. Statistical significance determined by using two-way analysis of variance (panels B, C, M, and N) or unpaired t-test (panels A and D-K). ∗P < .05, ∗∗P < .01, ∗∗∗∗P < .0001. Ctrl, control; PMN, polymorphonuclear neutrophil.